太平洋牡蛎同化率的研究

在室内实验条件下,研究了体重、饵料密度和质量对太平洋牡蛎同化率的影响.结果表明,太平洋牡蛎的滤水率、摄食率随个体的增大而增加,并且都符合幂函数模式:FR(IR)=aW^b.太平洋牡蛎同化率与其个体大小的关系不明显.饵料丰度和质量是影响太平洋牡蛎同化率的重要因素.随食物中有机物含量(POM/TPM)的增加,太平洋牡蛎的同化率也增加,随饵料密度的增加而降低.     

太平洋牡蛎同化率的研究

在室内实验条件下,研究了体重、饵料密度和质量对太平洋特蛎同化率的影响。结果表明,太平洋牡蛎的滤水率、摄食率随个体的增大而增加,并且都符合幂函数模式：ＦＲ（ＩＲ）αＷ＾ｂ。太平洋牡蛎同化率与其个体大小的关系不明显。饵丰度和质量是影响太平洋牡蛎同化率的重要因素,随食物中有机和的含量（ＰＯＭ／ＴＰＭ）的啬太平洋牡蛎的同化率也增加,随铒料密度的增加而降低。     

Noradrenaline reduces the stimulatory effect of interleukin-1α on reactive oxygen species production by oyster immunocytes

Abstract. A growing body of evidence suggests that interleukin-1α (IL-1α) is present in invertebrates. Both invertebrate and human IL-1α can bind to invertebrate receptors and stimulate invertebrate immune functions. The present study shows that IL-1α increases reactive oxygen species (ROS) production by oyster immunocytes. However, physiological doses of noradrenaline (NA) exert a suppressive effect on IL-1α stimulation in vitro . The β-adrenoceptor agonist isoproterenol mimicked the effects of NA and the β-adrenoceptor antagonist propanolol blocked the NA-induced suppression of hemocyte responsiveness to IL-1α. The type IV phosphodiesterase inhibitor rolipram acted in synergy with isoproterenol to reduce hemocyte response to IL-1α and the protein kinase A inhibitor H-89 suppressed the effects of isoproterenol. These results suggest that circulating NA impairs IL-1α-stimulation of oyster hemocyte via a β-adrenoceptor/cyclic AMP/protein kinase-A signaling pathway. Considering that mollusc immunocytes secrete NA, an autocrine regulatory loop may also modulate the ability of these cells to respond to IL-1α.     

太平洋牡蛎酪氨酸酶基因家族的系统发生分析

文章利用生物信息学方法对太平洋牡蛎(Crassostrea gigas Thunberg)酪氨酸酶基因家族的氨基酸序列特征、分类及系统发生进行了分析。结果表明, 太平洋牡蛎酪氨酸酶基因家族在进化过程中存在基因扩张现象, 其主要方式是基因重复。太平洋牡蛎酪氨酸酶可分为3种类型：分泌型 (Type A), 胞内型 (Type B)和具跨膜结构域型 (Type C)。根据太平洋牡蛎酪氨酸酶进化树分析, 发现Type A酪氨酸酶中, tyr18与其他Type A酪氨酸酶分化较大, 可能是较早分化出来的酪氨酸酶; Type B酪氨酸中的tyr2和tyr9以及Type C中的tyr8为较早分化出的酪氨酸酶。系统发生树分析发现太平洋牡蛎酪氨酸酶的聚类受酪氨酸酶类型以及基因位置的影响, 其分泌型酪氨酸酶首先与头足类分泌型酪氨酸酶聚在一起, 然后与线形动物门分泌型酪氨酸酶聚在一起, 与腔肠动物门分泌型酪氨酸酶分化明显。太平洋牡蛎胞内型酪氨酸酶自身分化较大, 总体上与线性动物门、其他软体动物胞内型酪氨酸酶聚为一支, 与扁形动物门、脊索动物门、腔肠动物门胞内型酪氨酸酶分化较大。太平洋牡蛎具跨膜结构域型酪氨酸酶与扁形动物门、环形动物门以及脊索动物门具跨膜结构域型酪氨酸酶分化明显, 与合浦珠母贝具跨膜结构域型酪氨酸酶聚为一支。这表明双壳类的Type C型酪氨酸酶与其他物种的同源酶的进化差异较大。文章首次探讨了太平洋牡蛎酪氨酸酶家族分类、分化及系统发生, 以期对太平洋牡蛎酪氨酸酶基因家族的理论研究和实际应用提供依据。     

Bioinformatic mining of EST-SSR loci in the Pacific oyster, Crassostrea gigas

A set of expressed sequence tag-simple sequence repeat (EST-SSR) markers of the Pacific oyster, Crassostrea gigas, was developed through bioinformatic mining of the GenBank public database. As of June 30, 2007, a total of 5132 EST sequences from GenBank were downloaded and screened for di-, tri- and tetra-nucleotide repeats, with criteria set at a minimum of 5, 4 and 4 repeats for the three categories of SSRs respectively. Seventeen polymorphic microsatellite markers were characterized. Allele numbers ranged from 3 to 10, and the observed and expected heterozygosity values varied from 0.125 to 0.770 and from 0.113 to 0.732 respectively. Eleven loci were at Hardy-Weinberg equilibrium (HWE); the other six loci showed significant departure from HWE (P < 0.01), suggesting possible presence of null alleles. Pairwise check of linkage disequilibrium (LD) indicated that 11 of 136 pairs of loci showed significant LD (P < 0.01), likely due to HWE present in single markers. Cross-species amplification was examined for five other Crassostrea species and reasonable results were obtained, promising usefulness of these markers in oyster genetics.     

长牡蛎精子超低温冷冻后超微结构损伤研究

采用程序降温仪分步降温冷冻保存长牡蛎（Crassostrea gigas）精液,并用扫描电镜、透射电镜研究了精子的超微结构损伤。超低温冷冻保存后长牡蛎精子的运动率、受精率及孵化率与鲜精无显著差异。鲜精中84.5%的精子形态结构正常,冻精中73%的精子形态结构正常。形态结构正常的精子表现为顶体、质膜、线粒体与鞭毛结构完整、染色质形状规则,顶体、线粒体及中心粒结构正常,鞭毛形态完整、微管结构清晰;形态结构异常的精子表现为顶体脱落、解体,精子头部质膜膨胀、破裂、染色质肿胀、破裂、解体,线粒体移位、脱落、膨胀,嵴退化或消失,鞭毛弯折、断裂,微管解聚。结果显示,以10% DMSO为抗冻保护剂,HBSS溶液为稀释液,1:4的稀释比例,添加海藻糖,采用分步降温法冷冻保存,对长牡蛎精子具有较好的抗冻保护作用,合适的冻存方法可以有效的保护太平洋牡蛎精子冷冻过程中结构损伤。研究有助于长牡蛎种质资源的收集保存及应用。     

蒽对太平洋牡蛎不同发育时期抗氧化酶活性差异性影响

在实验条件下采用生态毒理学和生物化学方法,选用常见的环境污染物多环芳烃蒽(Anthracene),以太平洋牡蛎(Crassostrea gigas)为实验材料进行毒理实验。研究了太平洋牡蛎D型幼虫、壳顶幼虫、幼贝和成贝4个不同发育时期中的3种抗氧化酶:超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)对蒽胁迫的敏感性。结果表明,在蒽胁迫下,D型幼虫和壳顶幼虫期3种抗氧化酶敏感性均为:POD>SOD>CTA;在幼贝和成贝期,其敏感性为:SOD>CTA,而POD表现出不稳定性。     

Sporulation of Minchinia sp. (Haplosporida, Haplosporidiidae) in the Pacific Oyster Crassostrea gigas (Thunberg) from the Republic of Korea

SYNOPSIS. A haplosporidan parasite belonging to the genus Minchinia was found in 4 of 1,438 oysters, Crassostrea gigas , collected from the Republic of Korea. Multinucleated vegetative stages were present in the infected oysters. Spores found in one of these oysters were acid-fast and had operculate characteristics of the genus Minchinia .     

5-HT和GnRH及其受体在长牡蛎卵巢的生理作用机制——双染和免疫细胞化学定位

用免疫细胞化学与双染技术对5-HT和GnRH及其受体在长牡蛎卵巢中的分布的研究结果提示,不同发育时期卵巢中卵母细胞均存在5-HT和GnRH受体,受体阳性物质沿胞膜分布,显深棕色,核显免疫阴性反应;5-HT和GnRH及其共存于不同时期卵母细胞胞质和胞膜上。以上结果表明长牡蛎的卵母细胞不仅能产生5-HT和GnRH,而且能表达5-HT和GnRH的受体,5-HT和GnRH可能以自分泌的方式对卵母细胞起调节作用。     

太平洋牡蛎养殖与野生群体遗传变异的微卫星研究

应用微卫星标记技术研究5个中国太平洋牡蛎养殖群体和2个日本太平洋牡蛎野生群体的遗传变异。研究中所使用的7个微卫星位点在养殖和野生群体中都显示出了高多态性,平均等位基因数为19.1～29.9,平均期待杂合度为0.916～0.958。养殖群体和野生群体的平均等位基因丰度及观察杂合度没有显著性差异。遗传分化系数及等位基因杂合度分析显示所有的群体间都有显著性差异。构建的NJ树中,7个群体聚为3支,养殖群体和野生群体可以清楚地分开,在养殖群体中又分为南北两支。分配检验中,97%～100%的正确率证明了微卫星标记在群体识别分析中的可行性。本研究结果对太平洋牡蛎管理模式的设计和选择育种具有重要意义。     

Microsatellite Analysis of 6-Hour-Old Embryos Reveals No Preferential Intraspecific Fertilization Between Cupped Oysters Crassostrea gigas and Crassostrea angulata

Experimental examination of reproductive isolation is the first step in understanding hybridization processes. Here, we studied preferential fertilization between 2 cupped oyster taxa, Crassostrea angulata and Crassostrea gigas, as a potential prezygotic reproductive isolation. Early examination of sperm competition is now possible by molecular analysis of oyster embryos. This avoids the confounding effect of differential mortality during the larval stage. Six hundred embryos were sampled from 2 crosses. Three microsatellite loci were enough to determine without ambiguity the taxa of contributing sires of embryos. No evidence of preferential fertilization between gametes from the same taxa was shown. A significantly higher contribution of the C. gigas males was revealed with the C. angulata females, but not with the C. gigas females, which might suggest early heterosis or interaction differences between gametes. In the light of these results, natural hybridization between both taxa can be expected in cases of their geographical coexistence, as in the Southern European populations in which both taxa are in contact as a result of aquaculture development. Received May 6, 2000; accepted March 6, 2001.     

Assessment and comparison of the Marennes-Oléron Bay (France) and Carlingford Lough (Ireland) carrying capacity with ecosystem models

Based on the individual growth, food limitation,population renewal through seeding, and individualmarketable size, a theoretical model of the culturedspecies population dynamics was used to assess thecarrying capacity of an ecosystem. It gave adome-shape curve relating the annual production andthe standing stock under the assumption of individual growth limited by the available food inan ecosystem. It also showed the influence ofmortality rate and marketable size on this curve andwas introduced as a means to explore the globalproperties resulting from the interactions betweenthe ecophysiology of the reared species and theenvironment at the ecosystem level. In a second step, an ecosystem model was built to assess the carryingcapacity of Marennes-Oléron bay, the mostimportant shellfish culture site in France, with astanding stock of Crassostrea gigas around 100000tonnes fresh weight (FW) and an annual production of30000 tonnes FW. The ecosystem model focused on theoyster growth rate and considered the interactionbetween food availability, residence time of thewater, oyster ecophysiology and number ofindividuals. It included a spatial discretization ofthe bay (box design) based on a hydrodynamic model,and the nitrogen or carbon cycling betweenphytoplankton, cultured oysters, and detritus. Fromsimulations of the oyster growth with differentseeding values, a curve relating the total annualproduction and the standing stock was obtained. Thiscurve exhibited a dome shape with a maximumproduction corresponding to an optimum standingstock. The model predicted a maximum annualproduction of 45000 tonnes FW for a standing stockaround 115000 tonnes FW. The prediction confirmedsome results obtained empirically in the case ofMarennes-Oléron bay and the results of thetheoretical model. Results were compared with thoseobtained in Carlingford Lough (Ireland) using asimilar ecosystem model. Carlingford Lough is asmall intertidal bay where the same species iscultured at a reduced scale, with current biomassless than 500 tonnes FW. The model showed that thestanding stock can be increased from 200 tonnes FWto approximately 1500 tonnes FW before any decreaseof the production. This revised version was published online in August 2006 with corrections to the Cover Date.     

Histological Characterization and Glucose Incorporation into Glycogen of the Pacific Oyster Crassostrea gigas Storage Cells

In order to investigate glycogen metabolism in the oyster Crassostrea gigas, the distribution of storage cells in the whole animal was studied before histological and biochemical characterization. These cells were found mainly in the labial palps, the mantle, and gonadal area and also in gills and the digestive area. Storage cells from palps, mantle, and gonad presented the same morphological features and the same seasonal glycogen variations. Storage cells were isolated from the labial palps and the mantle plus gonadal area of the oyster by enzymatic dispersion and centrifugation through discontinuous Percoll gradient. These cells have a modal density of 1.043 g/ml. An ultrastructural study confirmed that glycogen is present in the cytoplasm either as fine particles or sequestered within vesicles. Glucose incorporation into glycogen was evaluated in vitro using [U-¹⁴C]glucose: the incorporation in isolated cells increased linearly for at least 8 hours, was proportional to the cell concentration, and showed saturation kinetics with respect to the exogenous glucose concentration. Received March 18, 1999; accepted September 27, 1999.     

Identification and characterization of 18 novel polymorphic microsatellite makers derived from expressed sequence tags in the Pacific oyster Crassostrea gigas

We report the development of 18 new polymorphic microsatellite DNA markers derived from Crassostrea gigas expressed sequences tags. Genotyping of 48 wild adult oysters sampled from Marennes-Oléron bay (France) revealed 12 to 48 alleles per locus. Observed and expected heterozygosity levels ranged from 0.64 to 1 and from 0.77 to 0.97, respectively. The development of these new markers creates a useful complementary tool for population genetics studies, parentage analysis and mapping in Pacific oyster, a species of major aquacultural and ecological importance.     

Spatio-temporal localization of membrane lipid rafts in mouse oocytes and cleaving preimplantation embryos

We report for the first time the detection of membrane lipid rafts in mouse oocytes and cleaving preimplantation embryos. Cholera toxin β (CTβ), which binds to the raft-enriched ganglioside GM1, was selected to label rafts. In a novel application a Qdot reagent was used to detect CTβ labeling. This is the first reported use of nanocrystals in mammalian embryo imaging. Comparative membrane labeling with CTβ and lipophilic membrane dyes containing saturated or unsaturated aliphatic tails showed that the detection of GM1 in mouse oocytes and embryo membranes was consistent with the identification of cholesterol- and sphingolipid-enriched rafts in the cell membrane. Distribution of the GM1 was compared with the known distribution of non-raft membrane components, and disruption of membrane rafts with detergents confirmed the cholesterol dependence of GM1 on lipid raft labeling. Complementary functional studies showed that cholesterol depletion using methyl-β-cyclodextrin inhibited preimplantation development in culture. Our results show that the membranes of the mouse oocyte and zygote are rich in lipid rafts, with heterogeneous and stage-dependent distribution. In dividing embryos, the rafts were clearly associated with the cleavage furrow. At the morula stage, rafts were also apically enriched in each blastomere. In blastocysts, rafts were detectable in the trophectoderm layer, but could not be detected in the inner cell mass without prior fixation and permeabilization of the embryo. Lipid rafts and their associated proteins are, therefore, spatio-temporally positioned to a play a critical role in preimplantation developmental events.     

QTL for resistance to summer mortality and OsHV‐1 load in the Pacific oyster (Crassostrea gigas)

Summer mortality is a phenomenon severely affecting the aquaculture production of the Pacific oyster (Crassostrea gigas). Although its causal factors are complex, resistance to mortality has been described as a highly heritable trait, and several pathogens including the virus Ostreid Herpes virus type 1 (OsHV‐1) have been associated with this phenomenon. A QTL analysis for survival of summer mortality and OsHV‐1 load, estimated using real‐time PCR, was performed using five F₂ full‐sib families resulting from a divergent selection experiment for resistance to summer mortality. A consensus linkage map was built using 29 SNPs and 51 microsatellite markers. Five significant QTL were identified and assigned to linkage groups V, VI, VII and IX. Analysis of single full‐sib families revealed differential QTL segregation between families. QTL for the two‐recorded traits presented very similar locations, highlighting the interest of further study of their respective genetic controls. These QTL show substantial genetic variation in resistance to summer mortality, and present new opportunities for selection for resistance to OsHV‐1.